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1. HPTLC-MS
NAME- SEEMA SAMNTA SINGHAR
PA &QA DEPARTMENT
LECTURER CPS MOHUDA

2. Principle
• HPTLC based on simple principle which is separation of compound from
sample by adsorption phenomena.
• The basic principle of mass spectrometry (MS) is to generate ions from
either inorganic or organic compounds by any suitable method, to
separate these ions by their mass-to-charge ratio (m/z) and to detect
them qualitatively and quantitatively by their respective m/z and
abundance
• It is versatile tech. for separation as well as identification of pharmaceutical and
phytochemical.
• Traditionally the separation was carried out TLC separation and separation was
removed to study/identify in mass spectroscopy.
• The use of HPTLC and combination with high resolution TOF mass spectroscopy
for the detection, identify and imaging resulting in increase in analytical
importance.

4. Instrumentation
*For this follow HPTLC and mass spectroscopy instrumentation.
HPTLC:
• Sample Preparation
• Selection of Chromatographiclayers
• Plates
• Pre washing
• Conditioning
• Sample Application
• Pre conditioning
• Mobile Phase
• Chromatographic Development
• Detection of spot
• Scanning and Documentation
(*SEE HPTLC NOTE)

5. Interfaces:
High performance thin layer chromatography (Camag, Muttanz, Switzerland) consisted of an
auto sample applicator Linomat V connected to a nitrogen cylinder, TLC scanner attached to a
PC running win-CATS software (version 1.4.4), TLC Visualizer, and Camag twin-trough chambers
were used in analysis. HPTLC-MS detailed examination was carried out by TLC-MS interface
using acetonitrile as eluting agent at a flow rate of 1 ml/min. It remove/take out circular zones
in the form of bands from the developed HPTLC plate. The eluted material was transferred
automatically to single-quadrupole mass spectrometer, and mass spectra was recorded.
Surveys have shown that not all samples may be processed by HPTLC-MS or HPTLC-DAD or
HPTLC-MALDI or low delectability of the compounds or impurities in the UV range, a heavy
matrix load or a deficiency of MS compatible solvents. On the other hand HPTLC is another
very fast and suitable method to separate samples. In the past unknown substances were
scraped off from the TLC/HPTLC plate, eluted into a tube with solvent and transferred into the
MS. Now a very suitable and universal TLC-MS Interface is available which can semi-
automatically extract zones of interest and direct them online into any brand of HPLC-MS
system. The interface is fast and simply connected (by two fittings) to any LC-coupled mass
spectrometer without adjustments or mass spectrometer modifications. Questioned materials
are directly extracted from a TLC/HPTLC plate and sensitive mass spectrometric signals are
obtained within a minute per substance zone. The interface extracts the complete material
zone with its depth profile and thus allows detections comparable to HPLC down to the
pg/zone range. The interface has been demonstrate to be one of the most reliable and
versatile interfaces for TLC/HPTLC-MS coupling.

6. Mass spectroscopy:
Ion source
High vacuum system
Mass analyzer
Ion collector data system
(*SEE GC-MS OR MS NOTE)
Advantage of HPTLC-MS
It is cost effective because the chromatography run in decoupled with the detection
step.

7. Advantage
HPTLC-MS is a strong additional detection possibility and allows the structural
confirmation of targeted analytes. It also allows to do structure elucidation by coupling
HPTLC to high-resolution MS.

8. CONCLUSION:
1. This above short report a simple hptlc-ms method for the the routine analysis of
pharmaceutical product/formulation.
2.Its play important role in the study of the drug metabolism, discovery of new drug
candidate and the analysis, identification and characterization of impurities and degradent in
drug substants.
3. With regard to the outcome of this study and recommendations for plate handling, special
plate forms are rational.
4.The hptlc-ms interface is versatile building block of multidimensional liquid
chromatographic system.
5. Lc-ms have proved to be an extremely sensitive and specific technique for the analysis of
pharmaceutical product.
Application:
1. The liquid chromatography–mass spectrometry study of degraded products gave the idea
about the fragmentation patterns of degraded products. The developed method can be used
for routine analysis of terizidone and its formulations.
2. Degradation product of Empagliflozin in alkaline condition was carried out and its
degradation product is successfully separated and isolated by HPTLC method. degradation
product was identified by using MS-MS technique.